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Interactions of yeast dynein with dynein light chain and dynactin: General implications for intrinsically disordered duplex scaffolds in multi-protein assemblies Public Deposited

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https://ir.library.oregonstate.edu/concern/articles/wh246x807

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  • Intrinsically disordered protein (IDP) duplexes composed of two IDP chains cross-linked by bivalent partner proteins form scaffolds for assembly of multiprotein complexes. The N-terminal domain of dynein intermediate chain (N-IC) is one such IDP that forms a bivalent scaffold with multiple dynein light chains including LC8, a hub protein that promotes duplex formation of diverse IDP partners. N-IC also binds a subunit of the dynein regulator, dynactin. Here we characterize interactions of a yeast ortholog of N-IC (N-Pac11) with yeast LC8 (Dyn2) or with the intermediate chain-binding subunit of yeast dynactin (Nip100). Residue level changes in Pac11 structure are monitored by NMR spectroscopy, and binding energetics are monitored by isothermal titration calorimetry (ITC). N-Pac11 is monomeric and primarily disordered except for a single α-helix (SAH) at the N terminus and a short nascent helix, LH, flanked by the two Dyn2 recognition motifs. Upon binding Dyn2, the only Pac11 residues making direct protein-protein interactions are in and immediately flanking the recognition motifs. Dyn2 binding also orders LH residues of Pac11. Upon binding Nip100, only Pac11 SAH residues make direct protein-protein interactions, but LH residues at a distant sequence position and L1 residues in an adjacent linker are also ordered. The long distance, ligand-dependent ordering of residues reveals new elements of dynamic structure within IDP linker regions.
  • Keywords: isothermal titration calorimetry (ITC), nuclear magnetic resonance (NMR), protein assembly, intrinsically disordered protein, dynein
  • Keywords: isothermal titration calorimetry (ITC), nuclear magnetic resonance (NMR), protein assembly, intrinsically disordered protein, dynein
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  • Jie, J., Löhr, F., & Barbar, E. (2015). Interactions of Yeast Dynein with Dynein Light Chain and Dynactin: General implications for intrinsically disordered duplex scaffolds in multi-protein assemblies. Journal of Biological Chemistry, 290(39), 23863-23874. doi:10.1074/jbc.M115.649715
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  • 290
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  • 39
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  • This work is supported by National Institutes of Health Grant GM 084276 to EB. We acknowledge the support of the protein core facility in the OSU Environmental Health Sciences Center (NIH/NIEHS00210), and access to the Research Infrastructure activity in the 7th Framework Programme of the EC (Project number: 261863, Bio-NMR) (Frankfurt, Germany).
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